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Related Experiment Video

Updated: Jan 19, 2026

Derivation of Hematopoietic Stem Cells from Murine Embryonic Stem Cells
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Targeted Mutations in the Mouse via Embryonic Stem Cells.

Marina Gertsenstein1, Joffrey Mianné2, Lydia Teboul2

  • 1The Centre for Phenogenomics (TCP), Toronto, ON, Canada. marina.gertsenstein@phenogenomics.ca.

Methods in Molecular Biology (Clifton, N.J.)
|September 13, 2019
PubMed
Summary

This study details genetic modification techniques for mouse embryonic stem (ES) cells, crucial for creating mutant mouse models. It covers standard gene targeting and CRISPR/Cas9 methods to enhance genome manipulation efficiency.

Keywords:
CRISPR associated protein (Cas9)Clustered regularly interspaced short palindromic repeats (CRISPR)ElectroporationEmbryonic stem (ES) cellsGene targetingMouse

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Area of Science:

  • Genetics
  • Molecular Biology
  • Mammalian Biology

Background:

  • Genetic modification of mouse embryonic stem (ES) cells is vital for studying gene function.
  • This technology facilitates the creation of numerous mutant mouse lines.
  • Understanding mammalian biology relies heavily on these genetic tools.

Purpose of the Study:

  • To describe ES cell culture and transfection techniques for genome manipulation.
  • To outline standard gene targeting and CRISPR/Cas9 methods.
  • To enhance the efficiency of homologous recombination in ES cells.

Main Methods:

  • ES cell culture and transfection protocols.
  • Standard gene targeting for genomic manipulation.
  • CRISPR/Cas9 system application for double-strand DNA breaks.

Main Results:

  • Successful manipulation of the ES cell genome.
  • Generation of targeted ES cell clones.
  • Improved homologous recombination efficiency using CRISPR/Cas9.

Conclusions:

  • Established methods for genetic modification of ES cells.
  • CRISPR/Cas9 enhances gene targeting efficiency.
  • These techniques are fundamental for creating mutant mouse models.